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Cited by 52Alcohol Consumption and Health EffectsLiver physiology and pathologyHepatocellular Carcinoma Treatment and Prognosis

Adult mouse model of early hepatocellular carcinoma promoted by alcoholic liver disease

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Ambade, Aditya, Satishchandran, Abhishek, Gyongyosi, Benedek, Lowe, Patrick, Szabo, Gyongyi·2016·World Journal of Gastroenterology
DOI 10.3748/wjg.v22.i16.4091PMC PMC4837428
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Abstract

AIM: To establish a mouse model of alcohol-driven hepatocellular carcinoma (HCC) that develops in livers with alcoholic liver disease (ALD). METHODS: Adult C57BL/6 male mice received multiple doses of chemical carcinogen diethyl nitrosamine (DEN) followed by 7 wk of 4% Lieber-DeCarli diet. Serum alanine aminotransferase (ALT), alpha fetoprotein (AFP) and liver Cyp2e1 were assessed. Expression of F4/80, CD68 for macrophages and Ly6G, MPO, E-selectin for neutrophils was measured. Macrophage polarization was determined by IL-1β/iNOS (M1) and Arg-1/IL-10/CD163/CD206 (M2) expression. Liver steatosis and fibrosis were measured by oil-red-O and Sirius red staining respectively. HCC development was monitored by magnetic resonance imaging, confirmed by histology. Cellular proliferation was assessed by proliferating cell nuclear antigen (PCNA). RESULTS: Alcohol-DEN mice showed higher ALTs than pair fed-DEN mice throughout the alcohol feeding without weight gain. Alcohol feeding resulted in increased ALT, liver steatosis and inflammation compared to pair-fed controls. Alcohol-DEN mice had reduced steatosis and increased fibrosis indicating advanced liver disease. Molecular characterization showed highest levels of both neutrophil and macrophage markers in alcohol-DEN livers. Importantly, M2 macrophages were predominantly higher in alcohol-DEN livers. Magnetic resonance imaging revealed increased numbers of intrahepatic cysts and liver histology confirmed the presence of early HCC in alcohol-DEN mice compared to all other groups. This correlated with increased serum alpha-fetoprotein, a marker of HCC, in alcohol-DEN mice. PCNA immunostaining revealed significantly increased hepatocyte proliferation in livers from alcohol-DEN compared to pair fed-DEN or alcohol-fed mice. CONCLUSION: We describe a new 12-wk HCC model in adult mice that develops in livers with alcoholic hepatitis and defines ALD as co-factor in HCC.

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  • In the tumor microenvironment, efferocytosis promotes an immunosuppressive environment by polarizing tumor-associated macrophages toward a tumor-promoting M2 phenotype.

    “While it helps clear apoptotic tumor cells, it also promotes an immunosuppressive environment by polarizing tumor-associated macrophages (TAMs) toward a tumor-promoting M2 phenotype.”

    From Macrophage efferocytosis mediated by the TP63-RAC2 pathway promotes immunosuppressive remodeling in esophageal cancer (2026) · cites this paper as [9]

    The discussion of TAMs (section labeled “ROS Steatosis Proliferation Inflammation”; page unavailable) describes them as M2-skewed and says they can suppress adaptive immunity and promote tumors. The paper does not say that efferocytosis clears apoptotic tumor cells or causes TAMs to polarize toward M2; its experimental results instead report mixed M1/M2 markers in the liver. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • The M2-like TAM phenotype supports tumor growth, angiogenesis, and immune evasion.

    “This phenotype supports tumor growth, angiogenesis, and immune evasion, creating a formidable barrier to effective antitumor immunity.”

    From Macrophage efferocytosis mediated by the TP63-RAC2 pathway promotes immunosuppressive remodeling in esophageal cancer (2026) · cites this paper as [9]

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Authors
Ambade, A., Satishchandran, A., Gyongyosi, B., Lowe, P., & Szabo, G.
Affiliations
University of Massachusetts Chan Medical School (US)
ORCID
Venue
World Journal of Gastroenterology
Publisher
Baishideng Publishing Group Inc.
Published
2016
Volume
22
Issue
16
Pages
4091
ISSN
1007-9327 (ISSN-L)
DOI
10.3748/wjg.v22.i16.4091
PMC
PMC4837428
OpenAlex
W2344751387
Cited by
52
References
52
Citations by year
2026: 2 · 2025: 5 · 2024: 7 · 2023: 5 · 2022: 3 · 2021: 10 · 2020: 6 · 2019: 3 · 2018: 6 · 2017: 4
FWCI
3.94
Type
journal article
Language
en
Open access
Licence
cc-by-nc
Locations
  • World Journal of Gastroenterology (publishedVersion, cc-by-nc)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
  • The Journal of the American Medical Association (JAMA) Network (American Medical Association) (submittedVersion, cc-by-nc)
Funders
  • University of Massachusetts
  • National Institutes of Health
  • Center for AIDS Research, University of Washington
  • University of Massachusetts Medical School
  • National Institute on Alcohol Abuse and Alcoholism (#AA011576, R01AA011576)
  • National Institute of General Medical Sciences (T32GM107000)
Topics
  • Health Sciences › Medicine › Pathology and Forensic Medicine › Alcohol Consumption and Health Effects
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
  • Health Sciences › Medicine › Hepatology › Hepatocellular Carcinoma Treatment and Prognosis
Keywords
hepatocellular carcinoma, alcoholic liver disease, diethylnitrosamine, mouse model, macrophage polarization, M2 macrophages, neutrophils, hepatic steatosis, liver fibrosis, alpha-fetoprotein, hepatocyte proliferation, alcoholic hepatitis, CYP2E1
MeSH terms
Alanine Transaminase, Ethanol, alpha-Fetoproteins, Animals, Diethylnitrosamine, Endotoxins, Fatty Liver, Alcoholic, Carcinoma, Hepatocellular, Immunohistochemistry, Liver Cirrhosis, Alcoholic, Liver Neoplasms, Experimental, Macrophage Activation, Macrophages, Magnetic Resonance Imaging, Male, Mice, Inbred C57BL, Neutrophils, Phenotype, Time Factors, Biomarkers, Tumor, Cytochrome P-450 CYP2E1, Neutrophil Infiltration, Hepatocytes, Cell Proliferation

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